The ACE Scientific Core will provide support for the specialized immunological assays used by the two ACE projects, the Pilot Project, and potential clinical trials. Human and clinical research depends on reliable assays of valuable and often un-repeatable samples. Carefully-standardized methods are essential to ensure ,the quality of the rich datasets that can be generated by methods such as polychromatic flow cytometry. Thus a central Core focusing on the assay protocols helps to maintain comparability between different investigators, even in different institutions. The ACE Core works closely with the Rochester Human Immunology Center (RHIC), which develops and distributes expertise in cutting-edge immunological technologies. The RHIC develops new methods, polishes existing methods, and produces robust Standard Operating Protocols (SOPs) for many immunological assays. Close interactions with the RHIC ensure that the ACE Core has immediate access to new and improved RHIC methods, and conversely, that the RHIC is fully aware of method development that would benefit the autoimmunity research of the ACE Core. The ACE Core has acquired expertise in the following areas: polychromatic flow cytometry, using 11- and 18-color flow cytometers; staining panels for analysis of T cell cytokines, B cell and Treg subpopulations; gating strategies for many-parameter flow cytometry data; Elispot, Fluorispot and Cellspot assays for human antigen-specific cytokine- and antibody-secreting T and B cells; Functional human Treg and B cell helper assays; Multiplexed Luminex bead assays for detection of cytokines and other proteins; and centralized processing of clinical specimens in GLP conditions. The Core will support Project 1 with the development of flow cytometry panels for analysis of B cell subpopulations and cytokine production; and the large-scale analysis of flow cytometry data. Project 2 will be supported with the development and refinement of flow cytometric analysis of Treg populations; continued development of Treg suppressor assays; and the analysis of cytokine expression in mouse and human autoimmune effector T cell populations. The Pilot Project will be supported by flow cytometry analysis and B cell Elispots. The Core is available to support ACE Clinical Trials according to well-validated SOPs in all of these areas.